Figure 1.
T cell proliferation in normal or delipidated medium.
(A) T cells isolated from human peripheral blood were loaded with CFSE and cultured in culture medium supplemented with 10% delipidated FBS (delipidated) or 10% FBS (normal) for 4 days with CD3/CD28 activation beads. After activation, CD4+ and CD8+ T cells proliferated less when cultured in 10% delipidated FBS medium. Experiments were done in duplicates from the first sample, triplicates from the second sample and quadruplicates from the third sample using peripheral blood from 3 different human samples, hence n = 9 in each group. (B) Similarly CD4+ or CD8+ T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cultured in medium supplemented with 10% FBS. N = 6 in each group of CD4+ cells and n = 4 in each group of CD8+ cells.
Figure 2.
T cell proliferation in normal or delipidated meidum using serum of the same lot.
(A) T cells isolated from naïve apoE-/- mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4. (B) T cells isolated form naïve wild type mice proliferated less when cultured in medium supplemented with 10% delipidated FBS when compared to those cutured in medium supplemented with 10% FBS of the same lot. N = 4.
Figure 3.
Lipid raft and cholesterol content in T cells cultured in normal or delipidated medium.
T cells isolated from naïve apoE-/- mice were cultured in culture medium supplemented with 10% delipidated FBS or 10% FBS medium for 24 hours with CD3/CD28 activation beads. More unesterified cholesterol contents (A) or lipid rafts (B) were observed in activated CD4+ and CD8+ T cells cultured in 10% FBS medium when compared to T cells cultured in 10% delipidated FBS medium. Experiments were repeated 4 times using T cells from 3 mice.
Figure 4.
Western blot for pZap70 and IL-10 ELISA.
(A) T cells isolated from naïve apoE-/- mice were cultured in 10% delipidated FBS medium or 10% FBS medium for 2 min with CD3/CD28 activation beads. Western blot analysis of the whole cell lysates revealed a higher expression of pZap-70 from T cells cultured in NM (medium supplemented with 10% FBS) when compared to that from T cells cultured in DM (medium supplemented with 10% delipidated FBS). Upper panel showed a representative Western blot and lower panel is the densitometric analysis from three experiments. N = 3 in each group. (B) IL-10 level was lower in delipidated medium of T cells after activation for 4 days with CD3/CD28 beads when compared to that in normal medium. N = 4.
Figure 5.
Effects of diet change on atherosclerotic plaques in aortic sinus from apoE-/- mice.
(A) Representative photograph of Oil-Red-O and macrophage (MOMA-2) staining of atherosclerotic plaques in the aortic sinus of atherogenic diet (AD) and normal diet (ND) mice. Bar = 0.1 mm. (B) Representative photograph of CD3+ T cell staining of atherosclerotic plaques in the aortic sinus of atherogenic diet (AD) and normal diet (ND) mice. Bar = 0.1 mm. (C) Histomorphometric measurements of each staining are shown in bar graphs. N = 10 in each group for plaque size, lipid content and macrophage content. N = 5 in each group for CD3+ T cell content.
Table 1.
Effects of diet change on apoE-/- mice.
Figure 6.
Effect of diet change on IL-10 and IL-12 in T cells and their correlation with serum cholesterol.
(A) Flow cytometric analysis of splenocytes revealed that CD4+ and CD8+ T cells from AD mice express higher level of intracellular IL-10 and IL-12 when compared to those from ND mice. (B) Positive correlation was observed between serum cholesterol and the IL-10 positive or IL-12 positive in CD4+ or CD8+ T cells. N = 10 in each group. Pearson test was used for correlation.